Abstract 4278: Spheroid growth in 3D alginate matrices

Abstract

Abstract The use of 3-dimensional (3D) matrices for cell culture is gaining popularity as a substitute for traditional 2D cell culture as it can approximate cell architecture and cell-cell contact found in tissues, organs and tumors. NovaMatrix-3D™ is an alginate-based cell culture system comprising an alginate foam matrix and an alginate immobilizing solution. In principle, cells are first suspended in a sodium alginate solution then the cell suspension is applied to the alginate foam. In situ gelation occurs when calcium ions are donated from the foam cross-linking the added alginate, effectively entrapping the cells within the pores throughout the foam. To show the utility of the NovaMatrix-3D™ culture system, various cell lines were first cultured in 2D then prepared as a suspension in 0.5 – 1% sodium alginate in cell culture medium. An aliquot of each cell suspension was added to γ-sterilized NovaMatrix-3DTM foams (12- or 24-well plate size). NHIK 3025 (cervix carcinoma), MCF7 (breast adenocarcinoma), ZR-75-1 (breast ductal carcinoma) and NIH:OVCAR-3 (ovarian adenocarcinoma) were used and single cells were immobilized in the alginate foams. Cell localization within the foam was visualized using confocal microscopy to identify fluorescently labeled (CellTrace™ CFSE) cells. Cell proliferation was measured by counting cells after de-gelling the foam using sodium citrate. In each case, cells were immobilized with or without RDG-coupled alginate to investigate the importance of the presence of cell attachment peptides within the alginate. For these cell lines, cell growth and multicellular spheroid formation was independent of the presence of RGD. Spheroids could also be removed intact by de-gelling the alginate matrix and further processed for histological staining. One example shows the selective staining of apoptotic cells within the spheroid. Use of alginate foams with concomitant in situ immobilization of cells results in a 3D cell culture model with the potential to approximate cell proliferation and architecture within tissues or tumors. The technology enables biomimetic approaches by varying e.g. matrix elasticity, gelling ions, attachment peptides and foam degradation making NovaMatrix-3D™ a versatile cell culture system. Portions of this work have been funded with the support from METOXIA project no.222741 under the 7th Research Framework Programme of the European Union. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4278. doi:10.1158/1538-7445.AM2011-4278